The putative O-linked N-acetylglucosamine transferase SPINDLY inhibits class I TCP proteolysis to promote sensitivity to cytokinin

Evyatar Steiner, Sivan Livne, Tammy Kobinson-Katz, Lior Tal, Oded Pri-Tal, Assaf Mosquna, Danuše Tarkowská, Bruno Mueller, Petr Tarkowski, David Weiss*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


Arabidopsis (Arabidopsis thaliana) SPINDLY (SPY) is a putative serine and threonine O-linked N-acetylglucosamine transferase (OGT). While SPY has been shown to suppress gibberellin signaling and to promote cytokinin (CK) responses, its catalytic OGT activity was never demonstrated and its effect on protein fate is not known. We previously showed that SPY interacts physically and functionally with TCP14 and TCP15 to promote CK responses. Here, we aimed to identify how SPY regulates TCP14/15 activities and how these TCPs promote CK responses. We show that SPY activity is required for TCP14 stability. Mutation in the putative OGT domain of SPY (spy-3) stimulated TCP14 proteolysis by the 26S proteasome, which was reversed by mutation in CULLIN1 (CUL1), suggesting a role for SKP, CUL1, F-box E3 ubiquitin ligase in TCP14 proteolysis. TCP14 proteolysis in spy-3 suppressed all TCP14 misexpression phenotypes, including the enhanced CK responses. The increased CK activity in TCP14/ 15-overexpressing flowers resulted from increased sensitivity to the hormone and not from higher CK levels. TCP15 overexpression enhanced the response of the CK-induced synthetic promoter pTCS to CK, suggesting that TCP14/15 affect early steps in CK signaling. We propose that posttranslational modification of TCP14/15 by SPY inhibits their proteolysis and that the accumulated proteins promote the activity of the CK phosphorelay cascade in developing Arabidopsis leaves and flowers.

Original languageAmerican English
Pages (from-to)1485-1494
Number of pages10
JournalPlant Physiology
Issue number2
StatePublished - Jun 2016

Bibliographical note

Funding Information:
This work was supported by the Israel Science Foundation (grant nos. 576–11 and 779/15 to D.W.) and by the National Program of Sustainability I, MEYS, Czech Republic (grant no. LO1204 to P.T. and D.T.).

Publisher Copyright:
© 2016 American Society of Plant Biologists. All rights reserved.


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